Download Molecular Virology: Tables of Antimicrobial Factors

http://www.latrobe.edu.au/microbiology/table1.html
Página 1 de 14
Molecular Virology:
Tables of Antimicrobial Factors and Microbial Contaminants in
Human Milk
Molecular Virology:
Tables of Antimicrobial Factors and Microbial Contaminants in Human Milk.................................... 1
Table 1: Antibacterial factors found in human milk.......................................................................... 1
Table 2: Antiviral factors found in human milk................................................................................ 3
Table 3: Antiparasite factors found in human milk........................................................................... 5
Table 4: Microbial contaminants or nucleic acid detected in human milk........................................ 6
Table 5: Isolated contaminants from expressed human milk that caused infection...........................9
Table 6: Contaminants in infant formula that caused infections *.................................................. 10
Table 6a: Contaminants in infant formula that caused infections in hospitals................................ 11
Table 7: Effect of heat treatment or storage on antimicrobial factors in human milk..................... 11
Dr. John T. May and the Molecular Virology Laboratory............................................................... 13
Table 1: Antibacterial factors found in human milk
"Many cultures have considered that human milk has special medicinal and nutritional properties ...
it is also used as a folk remedy for conjuntivitis ... This view is paralleled in the 18th Century
London Pharmacopoeia which says, 'breast milk is an emollient and cool, and cureth Red Eye
immediately.'"
- Singh, et al. (1981) J. Trop. Ped. 28: 35
Factor
Shown in vitro to be active against
Secretory IgA
E. coli (also pili, capsular antigens, CFA1) including enteropathogenic strains, C.
tetani, C. diphtheriae, K. pneumoniae, S. pyogenes, S. mutans, S. sanguins, S.
mitis, S. agalactiae (group B streptococci), S. salvarius, S. pneumoniae (also
capsular polysaccharides), C. burnetti, H. influenzae. H. pylori, S. flexneri, S.
boydii, S. sonnei, C. jejuni, N. meningitidis, B. pertussis, S. dysenteriae, C.
trachomatis, Salmonella (6 groups), S. minnesota, P. aeruginosa, L. innocua,
Campylobacter flagelin, Y. enterocolitica, S. flexneri virulence plasmid antigen,
C. diphtheriae toxin, E. coli enterotoxin, V. cholerae enterotoxin, C. difficile
toxins, H. influenzae capsule, S. aureus enterotoxin F, Candida albicans*,
Mycoplasma pneumoniae
IgG
E. coli, B. pertussis, H. influenzae type b, S. pneumoniae, S. agalactiae, N.
meningitidis, 14 pneumoccoccal capsular polysaccharides, V. cholerae
lipopolysaccharide, S. flexneri invasion plasmid-coded antigens, major opsonin
for S. aureus
IgM
V. cholerae lipopolysaccharide, E. coli, S. flexneri
IgD
E. coli
Analogues of epithelial cell
receptors (oligosaccharides and
sialylated oligosaccharides***)
S. pneumoniae, H. influenzae
http://www.latrobe.edu.au/microbiology/table1.html
Página 2 de 14
Bifidobacterium bifidum
growth factors
(oligosaccharides,
glycopeptides)
Other Bifidobacteria growth
factors (alpha-lactoglobulin,
lactoferrin, sialyllactose)
Enteric bacteria. Two infant Bifidobacteria species provide a lipophilic molecule
which kills S. typhimurium. B. bifidum produces Bifidocin B which kills Listeria.
B. longum produces protein BIF, which stops E. coli.
Carbohydrate
E. coli enterotoxin, E. coli, C. difficile toxin A
Cathelicidin (LL-37 peptide)
S. aureus, group A streptococcus, E. coli
Casein
H. influenzae
kappa-Casein**
H. pylori, S. pneumoniae, H. influenzae
Complement C1-C9
(mainly C3 and C4)
Killing of S. aureus in macrophages, E. coli (serum-sensitive)
ß-defensin-1 or -2 or
neutrophil-α-defensin-1
or α-defensin-5 or -6
E. coli, P. aeruginosa, (some Candida albicans *)
Factor binding proteins (zinc,
vitamin B12, folate)
Dependent E. coli
Free secretory component**
E. coli colonization factor antigen 1 (CFA I) and CFA II, C. difficile toxin A,
H. pylori, E. coli
Fucosylated oligosaccharides
E. coli heat stable enterotoxin, C. jejuni, E. coli
Ganglioside GM1
E. coli enterotoxin, V. cholerae toxin, C. jejuni enterotoxin, E. coli
Ganglioside GM3
E. coli
Glycolipid Gb3
S. dysenterae toxin, shigatoxin of shigella and E. coli
Glycoproteins (mannosylated)
E. coli, E. coli CFA11, fimbrae
Glycoproteins (receptorlike)+ oligosaccharides
V. cholerae
Glycoproteins (sialic acid
-containing or terminal
galactose)
E. coli (S-fimbrinated)
alpha-Lactalbumin (variant)
S. pneumoniae
Lactoferrin**
E. coli, E. coli/CFA1 or S-fimbriae, Candida albicans *, Candida krusei*,
Rhodotorula rubra*, H. influenzae, S. flexneri, Actinobacillus
actinomycetemcomitans
Lactoperoxidase
Streptococcus, Pseudomonas, E. coli, S. typhimurium
Lewis antigens
S. aureus, C. perfringens
Lipids
S. aureus, E. coli, S. epidermis, H. influenzae, S. agalactiae, L. monocytogenes,
N. gonorrhoeae, C. trachomatis, B. parapertusis heat-labile toxin, binds
Shigella-like toxin-1
Lysozyme
E. coli, Salmonella, M. lysodeikticus, S. aureus, P. fragi, growing Candida
albicans* and Aspergillus fumigatus*
Milk cells (80% macrophages,
15% neutrophils,
0.3% B and 4% T lymphocytes)
By phagocytosis and killing: E. coli, S. aureus, S. enteritidis
By sensitised lymphocytes: E. coli
By phagocytosis: Candida albicans*, E. coli
Lymphocyte stimulation: E. coli K antigen, tuberculin
Spontaneous monokines: simulated by lipopolysaccaride
Induced cytokines: PHA, PMA + ionomycin
Fibronectin helps in uptake by phagocytic cells.
Mucin (muc-1; milk fat
globulin membrane)
E. coli (S-fimbrinated)
Nonimmunoglobulin
(milk fat, proteins)
C. trachomatis, Y. enterocolitica
http://www.latrobe.edu.au/microbiology/table1.html
Página 3 de 14
Phosphatidylethanolamine
H. pylori
(Tri to penta) phosphorylated
beta-casein
H. influenzae
Sialyllactose
V. cholerae toxin, H. pylori
Sialyloligosaccharides
on sIgA(Fc)
E. coli (S-fimbrinated) adhesion
Soluble bacterial pattern
recognition receptor CD14
Bacteria (or LPS) activate this to induce immune response molecules from
intestinal cells
Sulphatide
(sulphogalactosylceramide)
S. typhimurium
Unidentified factors
S. aureus, B. pertussis, C. jejuni, E. coli, S. typhimurium, S. flexneri, S. sonnei,
V. cholerae, L. pomona, L. hyos, L. icterohaemorrhagiae, C. difficile toxin B, H.
pylori, C. trachomatis
Xanthine oxidase
(with added hypoxanthine)
E. coli, S. enteritidis
Factors found at low level in
human milk
Shown in vitro to be active against
CCL28 (CC-chemokine)
Candida albicans*, P. aeruginosa, S. mutans, S. pyogenes, S. aureus, K.
pneumonidae
Heparin
Chlamydia pneumoniae
RANTES (CC-chemokine)
E. coli, S. aureus, Candida albicans*, Cryptococcus neoformans*
Secretory leukocyte protease
inhibitor (antileukocyte
protease; SLPI)
E. coli, S. aureus, growing C. albicans* and A. fumigatus*
* Fungi
** Contain fucosylated oligosaccharides. Stomach pepsin releases potent antibacterial peptides.
*** One sialylated pentasaccharide (3'-sialyllactose-N-neotetraose; NE-1530) had no beneficial effect on otitis media in
phase-2 clinical trials
Human milk contains nearly a thousand different oligosaccharides (determined by MALDI-mass
spectrometry). Many have the potential to act as receptors for bacteria not listed in the table.
Concentration of milk components in Breastfeeding: unravelling the mysteries of mother's milk (requires
completion of free registration to Medscape)
Various combinations of lysozyme, lactoferrin and SLPI have synergistic effect against E. coli.
Based on a table from the Proceedings of Breast Milk and Special Care Nurseries: Problems and Opportunities
Conference. August 1995. Melbourne.
NB: A bibliography for this table is currently available.
Content Approved by: John T. May
Page maintained by: Craig Lighton
Last Updated: 31 January, 2005
Table 2: Antiviral factors found in human milk
http://www.latrobe.edu.au/microbiology/table1.html
Página 4 de 14
"A variety of distinct antiviral factors were found in human colostrum and milk'"
- Sabin and Fieldsteel (1962) Pediatrics 29: 105.
Factor
Shown in vitro to be active against
Secretory IgA
Polio types, 1,2,3*. Coxsackie types A9, B3, B5, echo types 6,9, Semliki Forest virus, Ross
River virus, rotavirus*, cytomegalovirus, reovirus type 3, rubella varicella-zoster virus,
rhinovirus, herpes simplex virus, mumps virus, influenza, respiratory syncytial virus, human
immunodeficiency virus, hepatitis C virus, hepatitis B virus, hepatitis E, measles, sin
nombre hantavirus, SARS virus, Norwark and noroviruses.
IgG
Rubella, cytomegalovirus, respiratory syncytial virus. rotavirus, human immunodeficiency
virus, Epstein-Barr virus, sin nombre hantavirus, West Nile virus.
IgM
Rubella, cytomegalovirus, respiratory syncytial virus, human immunodeficiency virus, sin
nombre hantavirus, West Nile virus.
Bifidobacterium bifidum**
Rotavirus (by increasing mucin)
Chondroitin sulphate (-like) Human immunodeficiency virus
α defensins (1-3)
Herpes simplex virus, vesticular stomatitis virus, cytomegalovirus, influenza, human
immunodefiency virus
ß-defensin 1 or
α-defensin-5
Adenovirus
Haemagglutinin inhibitors
Influenza, mumps.
Lactadherin (mucinassociated glycoprotein)
Rotavirus*
Histo-blood group
carbohydrates
Norwalk virus
Lactoferrin
Cytomegalovirus, human immunodeficiency virus and reverse transcriptase, respiratory
syncytial virus, herpes simplex virus type 1, herpes simplex virus type 2, hepatitis C,
hepatitis B, poliovirus type 1, adenovirus 2 and Friend retrovirus. Also binds to the virus
receptors, low density lipoprotein receptor, and heparin sulphate proteoglycans. Hepatitis
G***, rotavirus*** and Seoul hantavirus***
Lipid (unsaturated fatty
acids and monoglycerides)
Herpes simplex virus, Semliki Forest virus, influenza, dengue, Ross River virus, Japanese B
encephalitis virus, sindbis, West Nile, Sendai, Newcastle disease virus, human
immunodeficiency virus, respiratory syncytial virus, Junin virus, vesticular stomatitis virus,
cytomegalovirus, mumps, measles, rubella, parainfluenza viruses 1-4, coronavirus, bovine
enterovirus (C12), poliovirus (C18), African swine fever virus.
Lysozyme
Human immunodeficiency virus, ectromelia
alpha2-macroglobulin (like) Influenza haemagglutinin, parainfluenza haemagglutinin.
Milk cells
Induced gamma-interferon: virus, PHA, or PMA and ionomycin
Induced cytokine: herpes simplex virus, respiratory syncytial virus.
Lymphocyte stimulation: rubella, cytomegalovirus, herpes, measles, mumps, respiratory
syncytial virus, human immunodeficiency virus.
Non-immunoglobulin
macromolecules
Herpes simplex virus, vesicular stomatitis virus, Coxsackie B4, Semliki Forest virus, reovirus
3, poliotype 2, cytomegalovirus, respiratory syncytial virus, rotavirus*.
Neutrophil-derived αdefensin-1 (HNP-1)
Herpes simplex virus 1
Ribonuclease
Murine leukaemia, human immunodeficiency virus
Secretory leukocyte
protease inhibitor
Human immunodeficiency virus, sendai, influenza
Sialic acid-glycoproteins
Adenovirus 37
slgA + trypsin inhibitor
Rotavirus
http://www.latrobe.edu.au/microbiology/table1.html
Página 5 de 14
Soluble intracellular
adhesion molecule 1 (ICAM- Rhinoviruses (major-group) 3, 14, 54; Coxsackie A13
1)
Soluble vascular cell
adhesion molecule 1
(VCAM-1)
Encephalomyocarditis virus
Sulphatide
Human immunodeficiency virus
(sulphogalactosylceramide)
Vitamin A
Herpes simplex virus 2, simian virus 40, cytomegalovirus
Factors found at very low
levels in human milk
Shown in vitro to be active against
Prostaglandins E2, F2 alpha Parainfluenza 3, measles
Prostaglandins E1
Poliovirus, encephalomyocarditis virus, measles
Gangliosides GM1-3
Rotavirus, respiratory syncytial virus, adenovirus 37
Gangliosides GD1a, GT1b,
GQ1b
Sendai virus
Glycolipid Gb4
Human B19 parvovirus
Heparin
Cytomegalovirus, respiratory syncytial virus, dengue, adenovirus 2 and 5, human
herpesvirus 7 and 8, adeno-associated virus 2, hepatitis C
* In vivo protection also.
** Used with Streptococcus thermophilus. Lactobacillus casei GG has also been used alone.
*** Only bovine so far, but human is normally identical.
Cytomegalovirus growth in vitro can be enhanced by the milk factors prostaglandins E1 or E2 or F2-alpha,
sialyllactose or interleukin-8.
Rotavirus growth can be activated in vitro by fatty acids (C10, C16).
HIV growth in vitro can be enhanced by (pro)cathepsin D. Prostaglandin E2 or transforming growth factor β
can either enhance or inhibit HIV depending on cell types infected.
Antibodies to CCR5 or lewisX sugar motif in milk can bind to HIV receptors.
HTLV-1 growth and cell infection can be enhanced by prostaglandin E2 or growth increased by lactoferrin or
transforming growth factor-beta.
Based on a table from the Proceedings of Breast Milk and Special Care Nurseries: Problems and Opportunities
Conference. August 1995. Melbourne. Copyright J.T. May and Australian Lactation Consultants Association
(ACLA) - Victorian Branch, 1995.
NB: A bibliography for this table is currently available.
Content Approved by: John T. May
Page maintained by: Craig Lighton
Last Updated: 6 February, 2006
Table 3: Antiparasite factors found in human milk
http://www.latrobe.edu.au/microbiology/table1.html
Página 6 de 14
Shown in vitro to
be active against
Factor
Secretory IgA
Giardia lamblia (protozoa)
Entamoeba histolytica (protozoa)
Schistosoma mansoni (blood fluke)
Cryptosporidium (protozoa)
Toxoplasma gondii
Plasmodium falciparum (malaria)
IgG
Plasmodium falciparum
Gangliosides
Giardia lamblia, Giardia muris
Lipid (free fatty acids
and monoglycerides)
Giardia lamblia
Entamoeba histolytica
Trichomonas vaginalis (protozoa)
Eimeria tenella (animal coccidiosis)
Lactoferrin (or pepsin-generated lactoferricin) Giardia lamblia, Plasmodium falciparum
Unidentified
Trypanosoma brucei rhodesiense
Macrophages
Entamoeba histolytica
Based on a table from the Proceedings of Breast Milk and Special Care Nurseries: Problems and Opportunities
Conference. August 1995. Melbourne. Copyright J.T. May and Australian Lactation Consultants Association
(ACLA) - Victorian Branch, 1995. Updated October, 1998.
NB: A bibliography for this table is currently available.
Content Approved by: John T. May
Page maintained by: Craig Lighton
Last Updated: 5 July 2004
Table 4: Microbial contaminants or nucleic acid detected
in human milk
"Transmission through breast milk seems to be the reason for the rapid and common aquisition of
cytomegalovirus that occurs among breast-fed infants. ..., it must be viewed as a form of natural
immunization."
- Stagno, et al. (1980) New Eng. J. Med. 302: 1073
Contaminant
Number of Infections
Viruses
#
B-type (retrovirus-like particles) Nil
Cytomegalovirus (or virus DNA)
About two thirds of infants consuming cytomegalovirus containing milk excrete
virus after 3 weeks. Up to a half of CMV positive mothers have varying levels of
infectious virus in their milk for up to 3 months. Present in preterm and mature
milk, but low in colostrum. One death in an infant with an immunodeficiency
syndrome. About 40% of preterm infants can be infected from non-frozen CMVcontaining milk. Symptoms may be seen in a quarter to a half of these infected
preterm infants.
http://www.latrobe.edu.au/microbiology/table1.html
Página 7 de 14
Epstein-Barr virus DNA
(glandular fever)
No increased seroconversion (infection) in breast fed infants.
Hepatitis B surface antigen
(or virus DNA)
No increased seroconversion (infection) in breast fed infants.
Hepatitis C RNA
Three infants had symptoms after breastfeeding for 3 months, from symptomatic
mothers with high levels of virus. Others have found no infection from chronic
infected mothers. Infants with hepC RNA may spontaneously clear the virus and
not seroconvert. Present in nil to 20% of infected mothers' milk*
Hepatitis E (or RNA)
Milk is not a major source, transmitted during pregnancy.
Herpes simplex virus type 1 (or
DNA) [cold sores]
One infected by 6 days. Infects also from nipple lesions, but infants may also
infect mothers. HSV-1 and HSV-2 DNA has been detected in milk cells.
Human herpesvirus 6 DNA*
(febrile illness)
Transmitted prior to breast feeding in HIV-infected infants. Present in the milk
cells of HIV-infected mothers. Cell-free virus was rare.
Human herpesvirus 7 DNA
(febrile illness)
No increased seroconversion (infection) in breast fed infants.
Human immunodeficiency virus
type 1 (and 2)
(or provirus DNA or virus RNA;
p24 antigen)
At least one third of transmissions to breast-fed infants is through milk. Most
occur by 5-6 months of breast feeding. HIV RNA can be present in half of infected
mothers' milk. The HIV variant (RNA) free in milk can be different to the proviral
(DNA) in milk cells in some mothers.
Human T-lymphotropic virus
type 1 (or provirus DNA; p24
antigen) [causes adult T-cell
leukaemia]
Transmitted to a quarter of infants almost exclusively through milk (cells) after 6
months of breast-feeding, in restricted geographical areas; seroconversion of
infants occurs after 12-24 months
Human T-lymphotropic virus
type II provirus DNA
Transmission occurs through milk
Rubella virus
A quarter of infants seroconvert 4 weeks after consuming rubella (normal or
vaccine strains) containing milk. Two thirds of vaccinated mothers can excrete
virus in milk for up to 3 weeks.
Sin nombre (no name)
hantavirus RNA [pulmonary
syndrome]
Nil
Transfusion-transmission virus
(TTV) DNA [no associated
disease]
Can be present in the milk of half to three quarters of women who have TTV DNA
in their serum (40% of women) and possibly transmitted to infants before
breastfeeding begins, or most probably (after 6 weeks) by later contacts, as
strains can vary from the mother's strain. *
Varicella-zoster virus DNA
(chicken pox)
One? Not found in recently vaccinated mothers' milk.
West Nile virus RNA
##
One without symptoms. WNV infection of mother was probably during postpartum
transfusion.
Bacteria
Borrelia burgdorferi DNA (Lyme
?
disease)
Brucella melitensis
Rare
Burkholderia pseudomallei
(Melioidosis)
Two?
Candida albicans ***
?
Citrobacter freundii
?; detected during infection in neonatal unit.
Coxiella burnetti (Q fever)
?
Enterbacter aerogenes
?; detected during infection in neonatal unit
Klebsiella pneumoniae
?; detected during infection in neonatal unit.
Lactobacillus gasseri /
Enterococcus faecium
(avirulent)
None? Present in the areola and colonise the infant gut as lactic acid bacteria.
http://www.latrobe.edu.au/microbiology/table1.html
Leptospira australis
Rare
Listeria monocytogenes
One?
Página 8 de 14
Mycobacterium paratuberculosis ?
Mycobacterium tuberculosis
(TB)
Nil?
Salmonella kottbus
One; may grow in milk ducts.
Salmonella panama
One
Salmonella senftenberg
One death; rare growth in milk ducts
Salmonella typhimurium
Rare
Serratia marcescens
?; detected during infection in neonatal unit.
Staphylococci
Rare. S. aureus or skin bacteria can be found in milk of mothers with mastitis.
Staphylococcus aureus (Pantonvalentine leukocidin producer;
associated with chronic boils)
One (pleuropneumonia)
Staphylococcus aureus
enterotoxin F
- ; mother had toxic shock syndrome
Streptococcus agalactiae (Group
Rare, one death; grows in milk ducts.
B streptococci)
Parasites
Necator americanus (new world
hookworm)
?
Onchocerca volvulus antigens
(skin worm)
Immune suppression
Schistosoma mansoni antigens
(blood fluke)
Hypersensitive allergy
Strongyloides fulleborni
(threadworm)
?
Toxoplasma gondii
One?
Trichinella spiralis (tissue
worm)
?
Trypanosoma cruzi * (Changas'
disease)
?
Other
Creutzfeld-Jacob transmissible
agent **
-
Mycotoxins (aflatoxins,
ochratoxin)
?; fungal toxins from food mother has eaten
* Not detected in all studies
** Never confirmed (New Eng. J. Med. 327: 649; 1992)
*** Fungi
# Detection of virus nucleic acid (RNA or DNA) does not mean the virus is still intact and infectious.
## A related virus, Central European encephalitis, has infected people through goats milk.
Syphilis may come from breast lesions
HIV-1 was possibly transferred in pooled unpastuerised milk that was fed to a young child for a four week
period (up to 15% of donors could have been HIV positive). Estimates of the time before HIV infection starts
to occur through milk vary widely, from four months to less than one month (most after four-six weeks).
One study reported HIV transmission is higher in mixed fed infants than those exclusively breast or infant
formula fed infants. Another shows little difference in exclusively breast fed or mixed fed infants, both
were significantly higher than formula fed infants at both six weeks and six months.
http://www.latrobe.edu.au/microbiology/table1.html
Página 9 de 14
Infants daily intake through milk may be 100,000 infected cells (HIV-1 or HTLV-1) or 10,000 infectious virus
(CMV or rubella), but each can be up to 100-fold higher. CMV infections appear to be from cell-free virus.
Whether CMV transmission from a CMV-positive mother to pre-term infant occurs depends on the viral load
(CMV DNA) in the milk.
Virus infections of infants take at least 3 weeks of feeding. There is no evidence indicating that one feed of
infected milk would cause a virus infection. Bacterial infections which are rarer can be quicker from
untreated expressed milk, but usually take about 3 weeks of feeding; but can also be treated using
antibiotics.
Group B Streptococci >100,000 cfu/ml has been found in an asymptomatic mother.
No hepatitis G / GB virustype C RNA or human herpesvirus 8 (Kaposi sarcoma-associated herpesvirus) DNA
has been detected in human milk.
Based on a table from the Proceedings of Breast Milk and Special Care Nurseries: Problems and Opportunities
Conference. August 1995. Melbourne. Copyright J.T. May and Australian Lactation Consultants Association
(ACLA) - Victorian Branch, 1995.
NB: A bibliography for this table is currently available.
Content Approved by: John T. May
Page maintained by: Craig Lighton
Last Updated: 14 October, 2005
Table 5: Isolated contaminants from expressed human
milk that caused infection
Contaminant
Number of Infections
Bacteria
Acinetobacter sp.
two
Enterobacter cloacae
two
Escherichia coli
several
Klebsiella oxytoca
two
Klebsiella pneumoniae **
six (three from a single donor)
Klebsiella sp.
six
Pseudomonas aeruginosa
one death, several infections
Serratia marcescens **
several
Staphylococcus epidermidis
(coagulase-negative) *
several; two deaths (mother's milk transported to twins)
Staphylococcus aureus
(methicillin-resistant)
several; one death (transported from mother)
Salmonella kottbus *
seven
http://www.latrobe.edu.au/microbiology/table1.html
Página 10 de 14
* from a single donor
** can multiply at room temperature. K. pneumoniae and P. aeruginosa has cross-contaminated pasteurised
milk.
Low levels of skin bacteria are normally found in expressed milk, which is normally bacteriostatic, high
levels (S. epidermidis above) are rare. The most common skin bacteria are S. epidermidis and to a lesser
extent Streptococcus viridans. Some bacteria indicated above were also introduced from incompletely
sterilised breast pumps (Klebsiella ssp., S. marcescens, P.aeruginosa and E. cloacae).
Milk expressed to be used in milk banks must contain < 100,000 cfu/ml to be pasteurised or < 10,000 cfu/ml
raw. Both exclude pathogens, S. aureus (coagulase-positive), group B streptococci and coliforms. No
agreed-upon guidelines exist for collected or frozen milk for mother's own milk, but < 100,000 cfu/ml is
frequently used. Higher levels (1,000,000 cfu/ml) of Gram-negative bacilli can be associated with sepsis.
Some methicillin-resistant S. aureus can grow and produce enterotoxin in colostrum at 37°C.
Four infants have died when fed milk with either Acinetobacter sp., Klebsiella sp. or coagulase-negative
Staphylococcus present (>10,000 cfu/ml).
One outbreak of F. meningosepticum was not from milk, but was located on milk bottle stoppers and
'cleaned' teats, as well as the ward environment.
Based on a table from the Proceedings of Breast Feeding, The Natural Advantage Conference. October, 1997.
Sydney. Copyright J.T. May and Nursing Mothers' Association of Australia. November, 1997.
NB: A bibliography for this table is currently available.
Content Approved by: John T. May
Page maintained by: Craig Lighton
Last Updated: 15 April, 2005
Table 6: Contaminants in infant formula that caused
infections *
Contaminant
Number of Outbreaks
Bacteria
Clostridium botulinum **
one infection? (UK, 2001)
Enterobacter sakazakii
several (various countries)
Salmonella anatum
one (UK / Europe, 1996)
Salmonella bredeney
two (Australia, 1977; France / UK, 1988)
Salmonella ealing
one (UK, 1985)
Salmonella london
one (Korea, 2000)
Salmonella tennessee
one (USA / Canada, 1993)
Salmonella virchow
one (Spain, 1994)
* Not contaminated during preparation for use
** Present in opened container, strain variation in unopened container
Other milk powders have been a source of infection in infants and adults, with different Salmonella or
Staphylococcus.
Milk powder added to bottles for infants became a source of one Bacillus cereus outbreak.
http://www.latrobe.edu.au/microbiology/table1.html
Página 11 de 14
It has been suggested that the high levels of galactomannan in cow's milk formula may be able to be
detected in infants sera leading to false positives for invasive aspergillosis.
US FDA bacterial compliance in formula. Formula meeting FAO food code may not meet some countries'
food laws (which can be <1 coliform/gram in all tests).
Copyright J.T. May, 2004.
Table 6a: Contaminants in infant formula that caused
infections in hospitals
Contaminant
Number of Outbreaks
Citrobacter freundii
one
Enterobacter sakazakii*** and
Leuconostoc mesenteroides***
one
Enterobacter sakazakii ****
several
Escherichia coli
one
Salmonella isangi
one
Salmonella saintpaul
one
Serratia marcescens
one
*** Has been isolated from blenders. In 1984 one report indicated Enterobacter cloacae was present in a manufacturer's
bottled formula.
**** The latest recall was in 2004. Notes on care with preparation of formula.
Other bacterial contamination has been traced to milk kitchen sources.
Copyright J.T. May, 2004.
NB: A bibliography for these tables is currently available.
Content Approved by: John T. May
Page maintained by: Craig Lighton
Last Updated: 5 December, 2005
Table 7: Effect of heat treatment or storage on
antimicrobial factors in human milk
Percentage of Activity Remaining *
Heat
Treatment
(15 seconds)
Heat treatment
(30 minutes)
Refrigeration
(7 days)
Freezing
(3 months)
http://www.latrobe.edu.au/microbiology/table1.html
Secretory IgA
72°C **
Flash
Pasteurisation
62.5°C
"Holding
method"
Pasteurisation
85
70
IgM
0
IgG
70
Lactoferrin (Ironbinding capacity)
100
Complement C3
Página 12 de 14
56°C
85
4°C
100
-15°C
100
Decreased
95
Decreased
40
75
100
0
0
90
Milk cells
0
0
0
10
Lyzozyme
100
75
100
90
100
100
100 ***
Vitamin A
Lipases (generate
antimicrobial lipids)
3
0
75
50
Other factors ****
100
(oligosaccharide, etc.)
100
100
100
100
Bacteriostatic activity
(on added E. coli)
Some decrease
Some
decrease
No decrease
Decreases
at 1 month, 66% present
at 3 months.
Gone in a quarter of
samples in 24 hours, all
gone by 7 days
Gone in most samples
after 24 hours, others
decreased by 99% in 3
days.
Same
Decreased
Cytomegalovirus
Nil
Nil
Can be
some
Skin bacteria
99% gone
Nil
Nil
* Values indicated are maximum values
** Special equipment needed for this high temperature treatment
*** Minimum of 3 weeks
**** These survive over 80°C for >30 minutes, while other listed factors are totally destroyed
HIV is destroyed by milk pasteurisation. HIV-1 is reduced ten-fold at 56°C for 121 seconds and at 62.5°C for
10 seconds in liquid; hepatitis B is killed and hepatitis C almost eliminated in serum at 60°C for 10 hours;
parvovirus B19 (similar to TTV) is removed at 60°C for 3 hours or 30 minutes at 70°C in liquid.
HTLV-1 (all cell-associated) is destroyed within 20 minutes at 56°C (or 10 minutes at 90°C), or by freezing
at -20°C for 12 hours. Cell associated HIV provirus DNA is destroyed by bringing milk to the boil. Boiling
milk destroys the immunoglubulins, lactoferrin, lysozyme and the milk's bacteriostaic activity, but not the
peptide beta defensin-1.
Pretoria pasteurisation (J. Trop. Ped. 2000, 46: 219) has been devised in an attempt to kill HIV, by standing
milk (50-150ml) in a glass jar in 450ml of preboiled water. The milk temperatures can remain between 5662.5°C for 10-15 minutes. Similarly, single bottle pasteurisers are available where basically boiling water is
added to a thermos flask containing the milk in a plastic bottle. A temperature of 58°C is reached in five
minutes and held at 60°C for 30 minutes. A solar-powered device can also pasteurise HIV-infected milk at
60°C for 30 minutes (J. Soc. Gynacol. Inv. 2000, 7: 366). Rehandling of the pasteurised milk can
recontaminate it.
Mature milk stored at room temperature for up to 6 hours (27-32°C) does not normally have any increase in
bacterial counts. However, S. epidermidis may have proliferated in a warm environment during collection
and transport (see Table 5).
Normally milk is not stored at 4°C for more than 48 hours and heat treated milk is stored frozen.
http://www.latrobe.edu.au/microbiology/table1.html
Página 13 de 14
Pasteurisation should kill all parasites which are rarely found in breast milk. Pasteurising human milk with
T. cruzi trypomastigotes inactivates the parasites.
Reconstituted infant formula will rapidly grow V. cholerae, S. flexneri and S. entertidis at 30°C but not if
refrigerated.
Very LBW babies are fed from milk banks with fresh frozen unpasteurised milk from donors who are also
CMV-IgG negative
After pasteurisation, milk has been contaminated with Pseudomonas aeruginosa when bottles (even with
tight lids) were cooled in cold water containing the organism. Also, 14 infants had symptomatic infection
with four dying of P. aeruginosa that contaminated milk from a pasteuriser and bottle warmer during
thawing of milk. Klebsiella pneumoniae has also cross-contaminated pasteurised milk.
Based on a table in the Proceedings of Breast Feeding: The Natural Advantage Conference. October, 1997.
Sydney. © J.T. May and Nursing Mothers' Association of Australia. November, 1997.
NB: A bibliography for this table is currently available.
Content Approved by: John T. May
Page maintained by: Craig Lighton
Last Updated: 30 November, 2004
Copyright: © 1997 J.T. May and Australian Breastfeeding Association
Dr. John T. May and the Molecular Virology Laboratory
The laboratory, which has been operational for over twenty-eight years, is centred around
animal virology, including molecular, biological and epidemiological studies. Eighty-five
publications have been produced from these studies.
We investigate anti-microbial factors and microbial contaminants of human milk and
continue our interest in this area by writing updating reviews for paediatric journals on a
regular basis. We first described the anti-viral lipids in human milk which subsequently were found to be
anti-parasitic and anti-bacterial also.
The major area of study of viruses has centred on herpesviruses. Initial studies on genital herpesvirus
(herpes simplex virus type 2) and a simple test for infection with this virus was patented world wide in 1984
and used in many countries. Current work centres on bovine herpesvirus 2 and its genes with attempts to
produce vaccines and to use this virus as a bovine vector virus for other cattle pathogens.
Another virus occasionally studied is the retrovirus human T-lymphotropic virus type I (HTLV-1) which we
first reported in Australian aborigines in 1988.
http://www.latrobe.edu.au/microbiology/table1.html
Content approved by: John May
Page maintained by: Craig Lighton
Last updated: 10 December, 2002
Página 14 de 14